Use of 1M NaCl split skin in the indirect immunofluorescence of the linear IgA bullous dermatoses

Willsteed, Elizabeth; Bhogal, B.S.; Black, Martin M.; McKee, Phillip H.; Wojnarowska, Fenella

doi:10.1111/j.1600-0560.1990.tb00073.x

Cited by 69 publications

(25 citation statements)

References 11 publications

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“…These results confirm previous reports of heterogeneity of the LAD antigens 3 –5 , 8 , 10 , 15 , 16 , 19 , 23 , 24 , 35 . There are also many reports which, taken together, demonstrate the multiple nature of the antigens 6 , 7 , 9 , 12 –14 , 17 , 21 , 36 .…”

Section: Discussionsupporting

confidence: 91%

“…The target antigens in LAD are still poorly understood. Indirect and direct immunofluorescence with salt‐split and suction‐split skin has demonstrated that the autoantibodies bind to the epidermal side, the dermal side, or in a combined pattern 3 –5 . Immunoelectron microscopy has localized the antigen to the hemidesmosome, 6 –8 the lamina lucida, 8 –10 the lamina densa 10 , 11 and the anchoring fibrils 12 –14 .…”

mentioning

confidence: 99%

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A comparison of the expression of known basement membrane components with the linear IgA disease antigens using the novel substrate cylindroma

Wojnarowska

Allen

et al. 1999

Br J Dermatol

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Linear IgA disease (LAD) is characterized by IgA basement membrane zone autoantibodies. Immunofluorescence, immunoblotting and immunoelectron microscopy studies have established the complexity and heterogeneity of the target antigens. We have studied the expression of the LAD antigens by cylindroma, a benign epithelial tumour that secretes abundant basement membrane, using 57 LAD sera categorized by indirect immunofluorescence on intact and salt-split skin. The expression of known components of hemidesmosomes, the anchoring filaments, extracellular matrix and the anchoring fibrils could be differentiated and were compared with the expression of the LAD antigens. The results showed that the LAD sera bound to the cylindroma tumour in two distinctive patterns. Thirty-three sera were positive on cylindroma. Twenty-seven sera bound to the basement membrane around the tumour clusters and the islets within the clusters in a thin linear band that was occasionally discontinuous. This was similar to the pattern observed with antibodies to the hemidesmosome components, the alpha6beta4 integrin and the bullous pemphigoid antigens BP230 and BP180. This pattern was observed with sera that bound to the epidermal (11) and the dermal (3) aspects of split skin or were negative (11), and with one serum which bound only to intact skin. Seven sera, all binding to the dermal aspect of split skin, bound the tumour cluster basement membrane in a thick band that was identical in appearance to that seen with antibodies to collagen VII; however, binding to the islets was either identical to collagen VII or similar but differentiated with double staining. Some sera were negative on cylindroma. Using fluorescence overlay antigen mapping we demonstrated colocalization of some epidermal-associated LAD target antigens with known hemidesmosome proteins, and colocalization of some dermal-associated LAD target antigens with anchoring fibril components. The results using cylindroma as substrate suggest that LAD autoantibodies may react with three or more target antigens. We propose from the results of this study that the autoantibodies in LAD target multiple antigens associated with hemidesmosomes and anchoring fibrils.

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Section: Discussionsupporting

confidence: 91%

mentioning

confidence: 99%

A comparison of the expression of known basement membrane components with the linear IgA disease antigens using the novel substrate cylindroma

Wojnarowska

Allen

et al. 1999

Br J Dermatol

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“…Some LAD sera that bound to the epidermis with salt-split skin changed to a combined pat tern with suction blisters. This pattern has previously been described by us with suction blisters and prolonged incuba tion with salt-split skin [2,30], These results suggest that there are at least 2 antigens. The change from the epidermal to the combined pattern with different methods of splitting may indicate 2 antigens or 2 epitopes, as there are differ ences in ultrastructure after salt and suction splitting [31].…”

Section: Discussionsupporting

confidence: 62%

Linear IgA Disease: A Heterogeneous Disease

1994

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Data from our series of 70 patients support the hypothesis that linear IgA disease is a heterogeneous disease as regards clinical features, target antigens and immunogenetics. The clinical presentation varies as regards age of onset and severity of skin and mucosal involvement and scarring. The use of split skin and of the novel substrate cylindroma has identified 2 target antigens. The majority of patients have a target antigen associated with epidermal cells and possibly cylindroma hemidesmosomes. A minority have a dermal antigen which resembles collagen VII on cylindroma. There is an association with HLA-B8, -DR3, -Cw7 and with the linked rare tumour necrosis factor a allele, but this is not universal. These differences do not correlate with each other.

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“…On the other hand, anti-BMZ.antibodies were detected by indirect IF only on saline-split skin, or their titers were higher on saline-split skin than on intact skin. These findings indicate the superiority of saline-split skin for the detection of anti-BMZ antibodies, confirming previous work (1,15). It is likely that chemical splitting produces better exposure of epitopes for anti-BMZ antibodies.…”

Section: Discussionsupporting

confidence: 91%

Concomitant Occurrence of Circulating IgA Anti‐intercellular and Anti‐basement Membrane Zone Antibodies in Autoimmune Blistering Diseases: Immunofluorescence and Immunoblot Studies

Dmochowski

Hashimoto

Bhogal

et al. 1993

The Journal of Dermatology

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Recently, cases with circulating IgA anti-intercellular antibodies have been described. The objective of this study was to present immunofluorescence and immunoblot findings in three cases of bullous diseases with concomitant circulating IgA anti-intercellular and anti-basement membrane zone antibodies. Direct immunofluorescence, indirect immunofluorescence on intact and 1M NaCl-split skin, immunoblotting of epidermal extracts from dispase- and EDTA-separated (two different procedures) human skin, and immunoblotting of the bovine desmosome preparation were performed. All three cases had IgA anti-intercellular and anti-basement membrane zone antibodies. However, immunoblot results were divergent. Case 1 had antibodies against the 150 kD pemphigus foliaceus antigen (IgG), the 170 kD protein (IgG and IgA), and the 97 kD antigen (IgG and IgA). Case 2 had IgG antibodies reactive with the 230 kD and the 170 kD bullous pemphigoid antigens, while case 3 had IgA antibodies against the 97 kD antigen only. The results of immunofluorescence and immunoblot studies in our patients widen the spectrum of laboratory features in blistering skin diseases mediated, at least in part, by antibodies of the IgA class.

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Use of 1M NaCl split skin in the indirect immunofluorescence of the linear IgA bullous dermatoses

Cited by 69 publications

References 11 publications

A comparison of the expression of known basement membrane components with the linear IgA disease antigens using the novel substrate cylindroma

A comparison of the expression of known basement membrane components with the linear IgA disease antigens using the novel substrate cylindroma

Linear IgA Disease: A Heterogeneous Disease

Concomitant Occurrence of Circulating IgA Anti‐intercellular and Anti‐basement Membrane Zone Antibodies in Autoimmune Blistering Diseases: Immunofluorescence and Immunoblot Studies

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